DE60021077T2 - DEVICE AND METHOD FOR SAMPLING - Google Patents

Description

FIELD OF THE INVENTION

The The present invention is directed to apparatus and methods for sample delivery directed. More particularly, the invention is to devices and methods for automatic microdimensioned sample delivery to chemical Reagents and / or analytical devices directed.

BACKGROUND OF THE INVENTION

method for execution Of chemical reactions often require several steps in several Reaction vessels, including one comprehensive handling of reagents. These restrictions can to experimental error, contamination and a risk of Exposure to dangerous Lead substances to laboratory workers.

The US-A-4906344 discloses a thermal technique for volume fluid movement in capillary electrophoresis. US-A-4979365 discloses a electrothermally operated actuator.

analysis techniques typically require a high level of work and use of complicated devices. In addition, many include chemical Laboratory and industrial processes the use of relatively large volumes of Reagents and several laboratory instruments. For example, require typical oversized Immunoassays the use of pipettes, reaction tubes and Reaction chambers. See, e.g. Mattiasson et al., Proc. Int. Symp. on Enzyme-Labeled Immunoassay of Hormones and Drugs (Pal, S., Hersg., Walter de Gruyter, Berlin (1978), p.91). Such processes can be ignored the size of the reaction too require several steps. Accordingly, there is a possibility for one reduced accuracy due to the introduction of impurities, Volume inaccuracies and low reproducibility. These applications serious in which biological samples, such as e.g. immunoassays Polynucleotide amplifications or hybridizations.

Recently Efforts have been made to modernize chemical processes Reduce costs, increase accuracy and reaction yields to improve. For example, capillary electrophoresis techniques have been suggested for a resolution to increase in immunoassays. Various attempts have been made to use other common analytical techniques, such as. the polymerase chain reaction (PCR). For example, For example, U.S. Patent 5,273,907 reports one with PCR reagents preloaded capillary, which is used to send a sample to the Reagents for DNA amplification leave. Similar describes International Patent Publication WO93 / 22058 micro-sized device for execution from PCR. In this case, PCR reagents are from a first chamber by moving materials through channels in a microchip with one Sample mixed in a second chamber.

It There is a need in the art for methods and devices, the work, costs, biohazardous exposure and complexity, those with a micro-sized sample delivery for chemical Reactions and analysis techniques is reduced. The The present invention addresses this need.

SUMMARY OF THE INVENTION

It Now devices and methods have been developed to automatically a sample to a reaction vessel, an analyzer or a deliver any location where sample introduction or deposition is desired. Generally speaking, the invention relates to a thermally controlled microdimensional sample delivery system and a method for his Use. An embodiment The invention comprises an apparatus and a method for dispensing a sample to a channel where chemical reactions occur.

One A sample delivery system of the invention as defined by claim 1, generally includes a housing, one channel, e.g. a capillary, limited, and a temperature control unit in thermal Connection with the channel. The channel is closed at one end and contains an opening for the introduction a sample. The closed end is typically connected to the temperature control device. The temperature control device a thermoelectric heating device, such as. a Peltier element, for heating and cooling a thermally expandable Be fluids in the channel. The temperature control device can also be a temperature-controlled Include fluid which is in thermal communication with the channel.

A sample delivery system of the invention preferably comprises an array of independent channels so that multiple samples can be dispensed simultaneously. The channels are often capillaries which may have non-wettable surfaces. In another embodiment of the invention, the channels are capillaries having immobilized therein at least one chemical reagent. In a preferred embodiment, a reagent in a capillary is preloaded by immobilization eg by drying the reagent on the walls of the capillary. Reagents can too by immobilization in a plug of material such as cotton placed in the capillary. Reagents are typically immobilized on the capillary walls in one or more discrete locations.

A other embodiment the sample delivery system includes a second temperature control device. The second Temperature control device can be used for heating and / or cooling the sample and reagents are positioned in the capillary. The second temperature control device preferably comprises a first conduit for heating and a second conduit for heating Cool. The second temperature control device is therefore adjusted to the temperature in a discrete part the capillary, typically towards the open end of the Capillary, to control. That is, the second temperature control device should no temperature changes of the fluid or gas near the cause the capillary to close. To this end For example, a sample delivery system can also have an isolation partition in the capillary included to help with a sample volume in the capillary stationary to keep.

Accordingly, the local Temperature of a reaction between reagents and sample through a second temperature control device without moving the sample in the capillary or by moving the second Temperature control device controlled along the capillary. A sample delivery system of Invention may contain more than two temperature control devices. However, one can single temperature control device be used to both the whole capillary and discrete Bodies to warm and / or to cool. A sample delivery system of the invention may include more than two temperature controllers.

method of the invention as in the claims 7 and 10 defined, take care of the delivery of a sample to pre-arranged reagents in a channel, to an input port an analyzer or to another location where the sample or its reaction products required become. In a preferred embodiment, a heated Temperature control device in conjunction with a capillary, the gas in the capillary so that the volume occupied by the gas increases, thereby his pressure increases. This increase in volume and pressure forces gas through the capillary opening. The opening the capillary is then exposed to a sample, e.g. by the open End in a liquid Sample is submerged. When cooling contracts the volume of gas remaining in the channel and the pressure in the capillary decreases. As a result, an aliquot a sample drawn into the capillary to the volume cavity to fill, which is left behind by the contracting gas. If sufficient cooling The sample is drawn far enough into the capillary to contact with chemical Reagents that are arranged to make contact, if any.

Products the reaction, if any, can be removed from the capillary by placing the gas close to the capillary Finally heated in the capillary becomes. Alternatively, the drawn into the capillary sample from the Capillaries are removed without any reaction, and to another reaction site, capillary, analyzer or somewhere, where a sample deposit desired will be delivered.

In another preferred embodiment For example, a sample delivery system of the invention is used to prepare a sample into a sample analyzer such as the one commonly owned US-A-6375817, incorporated herein by reference becomes. A sample analyzer (or sample plugger) comprises In general, a structure that limits two channels that are under at any angle to form a connection (or "joint"). one of the channels is a sample introduction channel with an opening for the introduction a sample. The other channel includes a separation channel in which a Medium can be arranged, which can separate components, of which it is suspected that they are in the sample. The sample analyzer further comprises means for applying a first pressure differential on the channels so that a sample flows into the connection. Then moved a second pressure difference part of the sample in the separation channel for separation and / or analysis. With the appropriate parameters and Control of pressure differences can be a well-defined sample plug be formed.

consequently The present invention provides apparatus and methods fast, accurate, automatic dispensing of samples to an analytical instrument kit or to chemical reagents for carrying out chemical reactions ready. When used in conjunction with a sample analyzer, The above may be the reaction and subsequent analysis be a fully automatic process for a sample.

The Invention will become apparent upon consideration of the following drawings, description and claims better Understood.

DESCRIPTION OF THE DRAWINGS

1 Figure 3 is a schematic illustration of a cross-section of an array of sample delivery systems of the invention, each capillary having reagents disposed therein to carry out chemical reactions.

2 Figure 3 is a schematic illustration of a cross section of a preferred sample delivery system of the invention comprising a capillary, a first temperature control device, and a second temperature control device.

3 Figure 3 is a schematic illustration of a cross-section of an array of preferred sample delivery systems of the invention comprising microfabricated channels, each having an inner diameter that varies along its longitudinal axis.

4 Figure 3 is a schematic illustration of a cross-section of an array of preferred sample delivery systems of the invention, including capillaries connected to a first temperature control device and a second temperature control device.

5A - 5F Fig. 3 are schematic illustrations of a cross section of a sample delivery system of the invention during an application of the invention.

6 FIG. 12 is a schematic illustration of a sample analysis device (also referred to as a sample plug forming device) having a sample introduction channel and a separation channel. FIG.

7 Figure 1 illustrates a unitized sample delivery system and sample analysis apparatus for rapid automated sample preparation and analysis.

Same Reference numerals in respective drawn figures show corresponding Parts on.

DETAILED DESCRIPTION OF THE INVENTION

The The invention generally provides a sample delivery system for automatic Delivery of a sample of reagents to carry out chemical reactions and / or analysis tools for the analysis of microdimensioned Rehearse. As used herein, the term "sample" is intended to mean any source suspected of that it contains a component, which is to be detected or identified, or any one potentially reactive chemical entity. A sample can be "undiluted" or can be used with a suitable solvent dilute be. Currently preferred samples include, but are not limited to biological specimens that are believed to be they contain a component of interest. Samples for use in the claimed invention include body fluids, such as. Blood, serum, plasma, urine, cerebrospinal fluid, Saliva, sweat, Tears, Seeds, vaginal fluid, Amniotic fluid and ascites fluid.

Generally In other words, a sample delivery system of the invention includes a housing that a channel is limited, such as a capillary. The channel has one first end up, that one opening for the introduction a sample and a second end associated with may be a volume control device. Typically, one is End sealed, with a closed end of the channel is limited. An opening for the introduction a sample in the channel is preferably opposite to the end of the Channels in conjunction with the volume control device, but they can exist anywhere along the channel.

The Volume control device can generally speaking a pump, a Syringe, a pipette balloon, a thermal regulator or another Facility for changing the volume and / or pressure in the channel. If the sample delivery system has a closed end, is the volume control device preferably a temperature control device in thermal communication with the closed end of the canal. The temperature control unit expands and / or contracted, i. heated and / or cool a thermally expandable fluid that is near the closed end is arranged in the channel. A regulation of the temperature of the thermally expandable Fluids in the channel move an aliquot of liquid, which is often a Contains sample of interest, into the canal and out of the canal. This way a can defined sample amount moves a specific distance in the channel be, then later ejected from the channel become.

One Sample delivery system of the invention may also include one or more chemical Reagents or phrases of chemical reagents disposed therein or immovable are made. In these embodiments is a sample through the channel for subsequent chemical reaction delivered to the reagents. If the chemical reaction at a Temperature performed If there is no ambient temperature, a second temperature control device may be connected to the Channel can be connected to the appropriate reaction conditions deliver. After the reaction can the reaction products for further reaction and / or analysis ejected from the channel become.

Accordingly, a sample delivery system of the invention may be used to perform numerous types of chemical reactions and / or to facilitate analysis of samples. For example, a sample delivery system may be used in diagnostic applications, such as a blood test (eg, to identify blood components or to detect / identify DNA in the blood), Im munoassays (eg, to detect the presence of a specific antigen in a sample) and colorimetric or other assays (eg, radiochemical, chemiluminescent, or binding assays). A sample delivery system of the invention may be used in applications to detect toxins (eg, bacteria, alcohol, drugs, viruses, organisms, metals, abnormal levels of physiological chemicals, and the like) or other components in a sample (eg, a biological or environmental sample). In addition, a sample delivery system can be used in chemical synthesis (eg, in the manufacture of drugs, peptides, nucleotides). A sample delivery system may also be used in a variety of laboratory techniques including, but not limited to, peptide or nucleotide sequencing, amplification and / or modification; Enzyme screening; and receptor-ligand reaction screening, which may use, for example, an antibody-antigen reaction.

It It is understood that the following discussion and examples are intended to be a preferred thermally controlled sample delivery system of the invention which is a capillary as the channel and a gas as the expandable fluid is used. That e.g., as herein incorporated by reference Gas, it is a specific reference to a Type of thermally expandable fluid, and the use of the term "gas" is a representative Example of a preferred embodiment, used to illustrate the teachings of the invention. However, the same principles and concepts that come through this description will be taught in the same way to the use any volume control device, channel and / or expandable Apply fluid known to those skilled in the art.

Regarding 1 is an array of sample delivery systems 6 represented according to the invention. The array of sample delivery systems 6 includes several capillaries 10 passing through an array holder 12 being held. The capillary may be made of glass, silica (fused silica) or polymeric materials, either inorganic or organic, such as suitable plastics.

The Capillaries can disposable (i.e., for single use) or reusable be. The capillary surface may be wettable or non-wettable. In another embodiment become the capillary channels in the surface etched of a substrate or molded. Although a microchip is typically a flat one surface which has several extensions and / or may require connections from the chip are skilled in the art Easily known techniques and materials to make a functional Microchip to manufacture, to apply the invention.

Of the Diameter and length a capillary can vary widely to provide the necessary dimensions, to create the necessary total volume of the capillary. Typical reaction volumes of the invention are less than about 30 microliters (μL) but preferably smaller. The inner diameter of each capillary typically ranges from about 5 μm to about 1000 μm. The inner diameter a capillary is preferably between about 20 microns and 300 microns. Even though Dimensions for a substantially circular one Cross sectional area a capillary, similar cross-sectional areas are preferred for noncircular channels, such as. rectangular channels with a depth and a width. The open end of the capillary may have a smaller diameter than the rest of the capillary. The Capillary may also have an inner diameter that runs along its longitudinal axis varies several times to provide different "zones" along the capillary. Even though microdimensioned samples are preferred and described, this limits in no way the invention, since a large-sized sample delivery using the principles and concepts disclosed herein are, can be achieved.

Again with respect to 1 is one end of each of the capillaries 10 sealed, with a closed end of the capillary 13 is limited. The closed end of the capillary 13 is with a temperature control device 14 connected and is in thermal communication with it. However, the temperature control device needs 14 not the entire closed end 13 to include, as in 1 played. The temperature control device 14 may be any heating / cooling element, the gas in the capillaries 10 can heat or cool, for example, a thermoelectric heating device. The temperature control device 14 may also be or include a thermally controlled fluid, such as water or polyethylene glycol, which can circulate through a constant temperature bath.

Preferably, the temperature control device may reach a preselected temperature in a particular time frame. The temperature control device is preferably capable of maintaining the preselected temperature within a tolerance range of a particular reaction for the required period of time. Thus, the temperature control device may be any suitable commercially available or custom made heating or cooling device that can achieve temperatures required to expand and contract the thermally expandable fluid in a channel. This particular thermally expandable fluid used to practice the invention prescribes the necessary temperatures that the temperature control device must reach.

One Temperature control device may also be an auxiliary control device, such. any suitable one microprocessor-based programmable logic controller, personal computer controller or the like for process control. A suitable control device includes features such as e.g. Programmability, reliability, flexibility and durability. The suitable control device comprises various Input / output ports used to provide connections around the temperature control unit to regulate, as well as to open valves and close Regulate and measure fluids, among other characteristics. The control device Also includes appropriate memory to process instructions for a desired application save. Naturally depends on that Controller type used by the specific application from.

A sample delivery system of the invention may be an insulating partition 15 include, as in 1 shown. The insulated partition 15 essentially separates the thermally expandable fluid in the vicinity of the closed end of the capillary 13 from the "reaction zone", which is the area in the capillary between the insulating partition wall 15 and the open end of the capillary 8th is. The term "reaction zone" refers generally to those portions of the capillary where reaction reagents are present, reactions are conducted, and reaction solvents or mixtures come into contact. In preferred embodiments, where the expandable fluid is a gas, the insulating partition is gas permeable. The insulating partition may be stationary or movable and rigid or flexible, depending on the construction materials for the partition and the particular application.

A Use of an insulated partition helps to achieve this thermally expandable fluid in the vicinity the closed end of the capillary at a temperature independent of Rest hold the capillary, allowing a movement of the sample in the Capillary essentially controlled by the temperature control device becomes. In this way, a reaction can take place at almost any temperature be carried out at a discrete point of the capillary, without the volume of reaction liquid due to expansion or contraction of the thermally expandable Fluids is moved.

One Isolator can also be outside the capillary and are in thermal communication with her. The outside insulator may be any suitable material that a high thermal conductivity that has for the special application is suitable. The outside insulator can also be in thermal communication with other materials, the derive a temperature. An external isolator can also be used in conjunction with an insulating partition.

Regarding 1 For example, a sample delivery system of the invention may further include one or more chemical reagents 16 contained in the inner walls of the capillaries 10 are arranged or immobilized on them. Immobilization of chemical reagents 16 can be achieved by drying on the inside of the capillary walls. The reagents 16 can pass through many procedures in the capillary 10 be dispensed, for example by being injected using a microneedle. After the chemical reagents 16 in the desired location in the capillary 10 can find the reagents 16 For example, by heating, dehydration or vacuum drying of the capillary 10 under appropriate conditions, dried so that the reagents 16 stay at their place of deposit. One or more of these drying techniques can be combined to form the reagents 16 to dry on the walls. The chemical reagents 16 can also be in the capillary 10 immobilized by being absorbed in a bulk material, such as cotton, placed in the capillary. In addition, certain chemical reagents 16 , such as PCR reagents, are dried in a suitable matrix, eg, dextran or trehalose, before being immobilized on the capillary walls.

More as a chemical reagent can in a capillary simultaneously to be available. That is, two or more chemical reagents may be present in the same Reaction zone present, creating a set of chemical reagents is defined. However, you can the two or more chemical reagents in the capillary are independent of each other spatially be spaced. It is understood that the use of the term "sentence" when describing chemical reagents one or more moieties or compounds of chemical reagents and not necessarily should mean more than one reagent.

Each set of chemical reagents is preferably dried in separate rings around the capillary walls. Relative placement of the reagents depends on many factors. That is, the dimensions of the capillary and the liquid volume of the reaction affect the placement of the reagents. In addition, the reaction conditions, including temperature and amount of reactants needed for reaction or analysis, affect the placement of the reagents in the capillary. In addition, the sequence of reactions to be performed affects space reagents, as contact of the sample with immobile reagents occurs sequentially as the sample in the capillary moves from the open end to the closed end, then back out the open end.

in the useful in the invention Reagents can be any chemical entity that is potentially one Probe or its component interacts. As the devices and Method of the invention in a wealth of chemical interactions useful are, the reagents useful in the invention are only by limited knowledge of a person skilled in the art. Accordingly, reagents include e.g. a binding protein, a nucleic acid probe, a PNA probe, an enzyme, a substrate, a ligand, a receptor, an antibody and / or a Antigen. Reagents can additionally Buffers, stabilizers, surfactants, additives, excipients, carriers, haptens or other compatible molecules include, which promote a reaction with sample components or influence. Reagents can for detection with a detectable entity or label or be marked for property modification. Include preferred detection marks, but are not limited to fluorescent, chemiluminescent, radioactive, mass spectrometric and colorimetric labels. Preferred Property Modification Markers include, but are not limited to, charge modifier tags, which can change the electrophoretic mobility of a reagent and Bio-recognition or chemical recognition labels that have improved analytical selectivity promote.

probes which are useful in a sample delivery system may include a be any nucleotide-binding compound, e.g. a riboprobe, a polynucleotide or a PNA. Preferably, the probe is complementary to one Target sequence present in the sample. In certain embodiments can Probes also binding proteins or other synthetic constructs be. It is preferred that the probes be detectably labeled are. Preferred labels include radioisotropic, fluorescent or colorimetric labels, enzymatic labels and Molecular weight markers, as well as other useful markers, the experts are known.

A Particularly preferred probe is a peptide nucleic acid (PNA). PNAs are DNA mimics with a neutral polyamide backbone, on the the nucleic acid bases are attached in the same way as they are on the phosphate backbone the DNA are attached. See, e.g. Egholm, et al., Nature, 365: 566-568 (1993); Oerum, et al., Nucl. Acids Res., 23: 5332-36 (1993); Practical PNA: Identifying Point Mutations by PNA Directed PCR Clamping, PerSeptive Biosystems Volume 1, Issue 1 (1995). See also the PCT publications EP 92/01219, EP 92/01220 and US 92/10921, which are incorporated herein by reference be recorded. Peptide nucleic acid probes typically form more stable duplexes with DNA compared to DNA / DNA duplexes. In addition, because PNA / DNA complexes a higher one thermal melting point than the analog DNA / DNA duplexes, a use of PNA probes the reproducibility of blotting assays improve. Peptide nucleic acid synthons and oligomers are commercially available. (PerSeptive Biosystems, Inc., Framingham, MA).

As used herein, the term "detectable Unity "any suitable label, including, but not limited to, enzymes, Fluorophores, biotin, chromophores, radioisotopes, colored particles, electrochemical, chemically modifying or chemiluminescent units. A currently preferred detectable moiety is a fluorescent Unit. Usual fluorescent Units include: fluorescein, cyanine dyes, coumarins, phycoerythrin, Phycobiliproteins, dansyl chloride, Texas red and lanthanide complexes. Naturally are the derivatives of these compounds known to those skilled in the art are, as usual including fluorescent moieties.

property modification include, but are not limited to, charge modifier tags, which can change the electrophoretic mobility of a reagent, and the bio-recognition or chemical identification markers that a promote improved analytical selectivity. Preferred Property Modification Markers are mass modification labels, e.g. Mass characteristics. preferred Charge modifying labels are known in the art, e.g. see US Pat. 5,630,924, incorporated herein by reference becomes.

Regarding 2 another embodiment of the invention includes a second temperature control device 20 in thermal communication with a capillary 10 , The sample delivery system used in 2 is shown, also has a first temperature control device 14 ' on, with the closed end of the capillary 13 connected is. The second temperature control device 20 is typically closer to the open end of the capillary 8th as the first temperature control device 14 arranged. The illustrated second temperature control device 20 has two lines 22 and 24 for controlling the temperature of the capillary 10 in the discrete zone, where each line with the capillary 10 thermally connected.

For example, as in 2 represented, the line 24 positioned so that the temperature of the capillary is regulated where the chemical reagents 16 immobilized. The invention can be practiced by the second temperature control device 20 is held stationary and the first temperature control device is used to move the sample or reaction mixture to the zone (or zones) in communication with the lines 22 and 24 to move. In these embodiments, one of the first conduit 22 or second line 24 maintain a first temperature, and the other of the lines can maintain a second temperature. However, in certain preferred embodiments, eg, in a PCR process, the reaction mixture preferably remains stationary in the capillary throughout the reaction process, ie, a thermal cycling occurs in the zone where the PCR reagents are immobilized. In these embodiments, the second temperature control device 20 stay stationary or can move along the longitudinal axis of the capillary 10 move.

In the first case, where the second temperature control device 20 remains stationary, is the second line 24 normally in thermal communication with the zone of the capillary 10 positioned where the chemical reagents 16 , as in 2 reproduced, are arranged. The second line 24 is used to control and maintain the temperature or temperatures required during a reaction or reactions, eg, thermal cycling. That is, a heated liquid or gas can pass through the second conduit 24 can be passed through to heat the reaction liquid volume, then a cooler liquid or gas through the second conduit 24 be passed through to cool the reaction mixture. In this way, a thermal cycling of the reaction mixture (also referred to as "reaction solution" or "reaction liquid volume") is achieved. Using this technique allows the first lead 22 as an insulator for the closed end of the capillary 13 acts, for example, by passing a fluid of constant temperature through the first conduit 22 flow. In other words, the first line 22 be used as an insulator to the thermally expandable fluid in the sealed end of the capillary 13 at a constant temperature, thereby ensuring that the reaction mixture remains substantially stationary throughout the reaction process.

In the second case, where the second temperature control device 20 along the longitudinal axis of the capillary 10 Anyone can move from the first lead 22 and second line 24 independently a different temperature in their respective zones of thermal connection with the capillary 10 maintained. As a result, the reaction mixture may remain stationary, and the second temperature control device 20 moves to a first temperature on the line 22 and a second temperature at the line 24 to deliver. It should be understood that although two lines are discussed and reproduced, a second temperature control device 20 The invention may include any number of conduits to provide the various temperatures and / or insulating features discussed above. In addition, a second temperature control device more than 2 Have lines.

3 Figure 12 illustrates a cross-sectional view of a preferred embodiment of the invention that is an array of microfabricated sample delivery systems. The channels are formed on a microfabricated solid, such as a glass or plastic substrate, which may be in the form of a microchip. The channels are typically etched or molded into the surface of the solid substrate, as discussed below. As in 3 reproduced, the channel indicates 10 ' an inner diameter that varies several times along its longitudinal axis. As a result, the channel has various zones (or regions) that allow isolation of different functions and / or responses.

For example, is in 3 the zone of channel 10 ' , which is referred to as letter "A", the thermal reserve area of the channel. This area includes the closed end of the canal 13 ' that is in thermal communication with a temperature control device 14 located. In the illustrated embodiment, the temperature control device is 14 a thermally controlled layer that is in thermal communication with the whole thermal control region "A" of the channel 13 ' located. As shown, the thermally controlled layer is behind the channels. However, the layer is preferably in front of the channels (or on top if the channels are horizontal). In addition, multiple layers can be used. The layer may be directly in thermal communication with the channels or may be in thermal communication with a substrate cover (not shown). Typically, the thermally-controlled layer is a heat sink coupled to a suitable heat controller that modulates the temperature of the fluid in the closed end of the channel.

The zone indicated by the letter "B" is a thermal insulation area where the channel 10 ' is constricted to help thermally isolate the thermal control region from the sample and / or the reagents. Zone "C" is the reagent mixture and incubation area or "reaction zone". In this area of the canal 10 ' can be a sample with chemical reagents 16 react that are optional. This Ge This may also be used to mix reagents and / or the sample that has been introduced to the channel via multiple introducer subchannels (not shown) or via a "bubble segregation" (discussed below). Zone "D" is the lead-in area where the sample and / or the reagents are delivered to the interior of the channel for transport therein. The channel in this area is often smaller in diameter than most of the channel. The channel in zone "D" may also include a plurality of open end and introduction subchannels converging at or near the reaction zone for mixing and the like therein.

microchips with channels can in large amounts from a solid substrate material designed and manufactured, which are easily sterilized can. Quartz glass is because of well-developed technology that its accurate and efficient fabrication enables a preferred substrate material, but other materials can can be used, including Polymers, e.g. Polytetrafluoroethylene. The channels can pass through a quartz glass substrate the most diverse machine micromachining processes, the Experts are known, inexpensive in big Quantities are manufactured. The machine micromachining process, the available Film deposition processes, such as e.g. spin coating and chemical vapor deposition, laser fabrication or photolithographic techniques, such as. UV or X-ray processes, or etching process, which are performed by either wet-chemical processes or plasma processes can. (See, e.g., Manz, et al., Trends in Analytical Chemistry 10: 144-149 (1991).) However, you can other manufacturing solutions be used.

Channels of varying Widths and depths can be manufactured with mikrodimensionierten dimensions. A machine Micromachining also provides a simple means to enable that a plurality of channels be manufactured and in mutual relationship, i. in fluid communication with each other to be brought. It should be apparent that a channel of Invention no consistent longitudinal axis like to have. That is, a channel may have multiple open ends, to the introduction of allow multiple samples and / or reagents to the channel. additionally a channel of the invention may have a closed end, which is in conjunction with several other channels, creating a common provided in a closed end, in conjunction with a Temperature control device stands, which simultaneously promotes movement of a liquid in the channels. Accordingly, many are different constructions and layouts of channels possible, depending on the specific application. Naturally apply the same design principles for capillaries. However, will complicated constructions are preferably micromachined by machine.

The channels may be enclosed using techniques known in the art. For example, the channels may be enclosed by bonding another planar substrate over the etched or embossed side of the microfabricated substrate. When the enclosed substrate is thin, it transfers heat quickly even if it is made of a thermally insulating material. Preferred thin film materials for enclosing microfabricated features include, but are not limited to, polyimide (eg Kapton ^®), Mylar (eg MonoKote), polyethylene, Teflon, glass, and laminates and composites of these materials. If desired for the application, reagents can be deposited on the walls of the channels and dried before enclosing the channels.

The Width and depth of a microfabricated channel can be set to promote functions, such as. Solution mixing, Lösungsabsondern, thermal insulation and interchannel manifold formation. Additionally, microfabricated multilayer sample delivery systems the functionality the sample delivery system by integration of other channel structures or integrated electronic devices known by those skilled in the art Increase procedure. Registration features can also imprinted in the substrate be used to promote assembly and robotic handling functions.

In addition, can the quartz glass substrate, containing machined channels, with a thin anodic bond Covered and sealed glass cover. Other transparent or opaque cover materials can be used. alternative can two microfabricated quartz glass substrates sandwiched or a quartz glass substrate can be placed between two glass covers added become. The use of a transparent cover leads to a Windows, which promotes a dynamic viewing of the channel contents and optical sampling of the channels either visually or through a machine allows.

The use of microfabricated channels in a solid state substrate as described above provides many advantages. The dimensions and shape of the channels can be adjusted for processes or functions that are not easily met using capillaries of a single diameter. For example, solutions drawn into a channel which are initially separated by a bubble may be mixed as the solutions are transported to a region of the channel having an increased diameter has. Alternatively, channels with reduced diameter ranges can be fabricated to reduce the heat transfer coefficient from one portion of the channel to another.

Additionally allow microfabricated channels, that a reagent solution is easily deposited in an area of an unclosed channel and subsequently is dried to leave the reagent in the channel before the channel with an enclosing Substrate is enclosed. Furthermore can multilayer microfabricated structures are formed, the the functionality the sample delivery system by channel manifold formation or by integration of electrical components in the sample delivery system using skilled persons increase known methods. In other embodiments can Registration features are etched or molded onto the substrate to an alignment of the substrate with other instrument components or other layers of the sample delivery system, whereby the alignment and handling of the substrates in an automatic Robot system is simplified. All the above features bear to reduce the complexity of the delivery system of the invention, by a single solid-state delivery system which contains an array of channels that are light and effective can be handled.

4 represents an array of capillaries 10 each representing a closed end 13 and in each case in conjunction with a first temperature control device 14 ' are located. The capillaries 10 each contain a first set of chemical reagents 18 and a second set of chemical reagents 26 immobilized on the capillary walls. A second temperature control device 20 that the wires 22 and 24 respectively for heating and / or cooling, allows control of the temperature in the discrete part of the capillary containing the chemical reagents, as discussed above. Use of the second temperature control device and an insulator (eg, an insulating partition, not shown) allows the temperature of the reactions to be controlled without thermally affecting the gas in the closed end of the capillary.

In In its broadest aspect, the methods of the invention are one Directed use of a sample delivery system described above to enter an aliquot of a sample to a desired location. The volume of fluid in a capillary is determined by the temperature of the fluid in the capillary is modulated. Preferably, the fluid is a Gas. However, the fluid may be an inert liquid. Regardless on it is the volume and pressure of the thermally expandable Fluids the agent that is used to sample into the capillary and move out of it.

The 5A - 5F disclose the various stages of a sample delivery system during the application of a method of the invention. 5A FIG. 3 illustrates a sample delivery system of the invention prior to use and similar to the system disclosed in FIG 1 is reproduced. Changing the temperature of a thermally expandable gas in the capillary causes gas to expand or contract. Typically, when a gas is heated, the volume it occupies increases, and hence the pressure in the capillary is approximately equal to the ideal gas law PV = nRT, where P is the pressure, V is the volume, n is the number of gas molecules , R is the constant 8.314 JK ^-1 mol ^-1 and T is the temperature in degrees Kelvin. When the gas passes through the temperature control unit 14 is heated and sealed one end of the capillary 13 is, the expanding gas ("G") escapes through an opening in the capillary 8th , following the path of least resistance, as in 5B shown. Accordingly, the pressure in the capillary 10 lowered. Subsequently, the capillary 10 in a sample 58 ( 5C ) and the gas is cooled. The pressure difference between the outside and the inside of the capillary 10 forces an aliquot of a sample 60 into the capillary 10 , That means the pressure outside the capillary 10 indicated by the letter "P" in 5C is shown, "presses" on the sample 58 to the pressure inside and outside the capillary 10 to balance. As in 5D shown, after cooling, an aliquot of the sample 60 into the capillary 10 introduced.

Preferably, the volume of a sample introduced into the capillary is a pre-selected or metered aliquot, typically by the time the capillary 10 in the sample 60 is submerged, and the pressure differences acting on the sample is determined. Subsequently, the aliquot of the sample 60 which is drawn into the capillary, deposited somewhere for further reaction and / or analysis (not shown).

Alternatively, the sample and reagent solutions may be introduced and / or metered by nonthermal means into a capillary. For example, an auxiliary volume control device in fluid communication with a capillary may be used to introduce a predetermined aliquot of a solution into the capillary. Another example is the use of capillary action to fill part of the capillary. With the proper construction, a predetermined amount of solution may be introduced into the capillary via capillary action to provide a metered aliquot. In addition, with a capillary having multiple open ends and introduction subchannels, multiple solutions may be co-extruded into a capillary tig and in measured quantities. Subsequent to entry of the solution into the capillary, thermally actuated volume changes in the capillary, as discussed above, may be used to transport the sample and / or reagents to other areas of the capillary.

In certain preferred embodiments, as in 5A - 5F reproduced, may be an aliquot of the sample 60 into the capillary 10 be pulled, leaving the aliquot of the sample 60 to a chemical reagent 16 is discharged and in contact with this, in the capillary 10 is preloaded. Upon contact of the aliquot of the sample 60 with the chemical reagent 16 if the conditions are suitable, a chemical reaction may occur, as in 5E shown. To promote a reaction or interaction, the reaction conditions may be changed using a second temperature control device (not shown), as described above, ie by heating or cooling that portion of the capillary where the reagent 16 is arranged. Subsequent to a reaction can reaction products 62 and other components, such as starting materials, from the capillary 10 be eluted to a suitable location by adjusting the temperature of the gas near the closed end of the capillary 13 is increased, as described above and in 5F played.

Alternatively, with the appropriate system and application, the reaction products 62 be moved to another zone of the capillary by placing the gas near the closed end of the capillary 13 using the temperature controller 14 is heated or cooled. In another zone, the reaction products 62 can be analyzed directly in the capillary or can contact a second chemical reagent for possible further reaction under the right conditions. It should be understood that if there is no reaction between the sample and the reagents, the sample may be moved and / or analyzed as above to provide useful information, the same as if a reaction had occurred.

A another technique using a sample delivery system of the invention can be applied is known as "bladder secretion". An aliquot a source sample is drawn into a capillary, the capillary is removed from the starting sample and then placed in a second sample. The second sample may be a solution of reagents. Upon further cooling of the gas in the Capillary nearby of the closed end becomes the second sample (or reagents) is drawn into the capillary and begins to move to mix with the starting sample at its interface. Depending on Many factors can control the mixing of the two samples. However, the introduction may the second sample into the capillary then make that a Volume of gas is first drawn into the capillary, causing a "bubble" between the first Sample and the second sample. This is "bladder secretion" because of a bubble the two liquids, which are drawn into the capillary, separating and preventing their mixing. Accordingly, on Basis of the aforementioned Techniques and others that are known to professionals, the most diverse useful Procedures are controlled and implemented to the special Requirements of an experimental protocol or application too correspond.

For example, as the solutions of separated bubbles (either samples and / or reagents) are transported to an area of the capillary where the capillary diameter increases, the bubble no longer forms an effective barrier between the two solutions and the solutions contact each other can mix. This application of a "bladder secretion" is in 3 played. As shown, became a first solution 21 to the open end of the channel 8th' introduced sequentially followed by a gas and a second solution 21 ' , The result is a gas bubble 23 that the first solution 21 from the second solution 21 ' secretes. Subsequently, as the solutions and the bubble move along the longitudinal axis of the channel away from the open end 8th' move into the reaction zone ("C") where the bubble is no longer effective to segregate the solutions, due to the increased volume of the channel 10 ' in this area. As a result, the solutions come in contact with each other and also with the chemical reagents 16 which are optionally arranged therein.

To assist in automating the methods described herein, another aspect of the invention is a scientific instrument incorporating the sample delivery systems described above. The scientific apparatus enables the efficient automation of the systems of the invention with their auxiliary equipment and equipment. The scientific device also allows other devices to be connected to the delivery systems of the invention to enable a functional design to meet end-user needs. For example, analytical instruments may be associated with a scientific instrument of the invention to enable analysis of samples, eg at given times in the reaction cycle. Analytical instruments useful in the invention are well known to those skilled in the art and include, but are not limited to, mass spectrometry instruments, chromatographic systems and various detection instruments, such as ultraviolet, infrared, fluorescence and refractive index detectors.

Other non-limiting Examples of auxiliary instruments, useful in the invention include diagnostic tools for performing assays and synthesizers to automate the production of special compounds, to become part of a sample. Such synthesizing devices include those that can perform combinatorial syntheses that screening libraries of compounds with the delivery systems enable the invention. All above instruments and devices can from Hand operated step by step. However, a full automation prefers. As for a person skilled in the art preferably includes automation a microprocessor and / or computer that has different aspects The method of the invention controls, but typically at least in connection with the temperature control unit.

To Existence of the disclosure of the basic operation and the basic principles, which are the basis of the invention, a person skilled in the art will recognize slightly different Sample Delivery and Chemical Reaction Schemes / Protocols Shown In Compound can be used with this invention. For example, can several Reactants may be present in the capillary, each by an inert Zone separated. The variation of reaction temperatures could be increased by several Temperature controllers arranged adjacent to each set of reactants are, or a second temperature controller can be controlled, the more Has lines that are positioned accordingly. consequently can the most diverse chemical reactions and processes through the sample delivery system promoted the invention or performed be inclusive but not limited on PCR.

One Sample delivery system of the invention can be used to provide a Sample, a reacted sample and / or other reaction products to a To submit device for analysis. A particularly preferred device is described in commonly owned US-A-6375817.

The The sample analyzer (or sample plugger) mentioned above has one casing on, the two channels limited, which intersect to form a connection, the promotes formation of a sample plug. Afterwards its formation becomes the sample plug along one of the channels, one Separation channel, to an analytical instrument and / or the separation of Sample transported before detection in their individual components.

As in 6 As shown, a first channel for introducing a sample is a sample introduction channel 27 , and a second channel, the sample introduction channel 27 is a separation channel 29 , The apparatus further comprises means for generating pressure differences in the channels, such as a vacuum pump or a peristaltic pump. The device may also include a voltage generator 43 for generating a voltage gradient along the separation channel 29 include. Finally, the device may comprise a detector for detecting components in the separation channel.

As As indicated above, in a preferred embodiment, the channels are on one microfabricated solids formed, such. a silica or quartz glass substrate, which may be in the form of a microchip. Each channel typically contains a suitable medium. The separation channel can be a medium for separation of sample components based on their charge or size. The medium may e.g. Sieve media, such as e.g. Polyacrylamide. However, you can other sieve media for a particular application, as will be apparent to one skilled in the art. Accordingly, a Sample dispensing system used in conjunction with a sample analyzer will, be used to a complicated reaction, separation and analysis protocols, e.g. Immunoassays or polynucleotide identifications perform.

An apparatus for analysis used in conjunction with a sample delivery system of the invention provides for the automatic uniform production of sample plugs through the use of vacuum and / or pressure on the sample introduction and separation channels. As in 6 illustrates the Probeneinführkanal 27 a connection 33 with the separation channel 29 , Applying pressure and / or vacuum to the sample introduction channel 27 , then the separation channel, causes a sample plug 35 downstream of the connection 33 in the separation channel 29 is formed. (It is understood that 6 is a schematic representation and that in reality the sample plug 35 contained in the channels.) Arrows 37 . 39 and 41 represent the direction of a sample flow. A voltage generator 43 if present, can apply a voltage gradient axially along the separation channel. A voltage gradient may be applied while a pressure gradient moves a sample along the sample introduction channel past the junction to apply one type of sample plug formation technique, referred to as a "cluster". Using a piling technique, a diluted sample can be separated before separation and / or analysis.

A sample delivery system of the invention, used in conjunction with a sample analysis apparatus described above, provides for the rapid, automatic analysis of biological samples without the complicated machinery, time, and biological hazard exposure inherent in the use of existing systems. 7 FIG. 12 illustrates a unitized sample analysis device and sample delivery system array. The unitized device 62 contains a sample card 28 with a platform or membrane on which a sample, such as blood, is deposited. The card may be, for example, an IsoCode ^™ card (Schleicher & Schuell, King, NH). The unit designed as a unit 62 further includes an array of sample delivery systems of the invention 6 , as described above. The device 62 contains a microchip assembly 32 with sample introduction and separation channels connected to a pressure / vacuum unit 34 , a high voltage power supply 36 and a high pressure cartridge 38 are connected.

Near the end of the separation channels of the microchip assembly is an optical scanning module 40 , The optical scanning module detects the presence of detectable moieties bound to the component of interest in the sample. Detection may be achieved by methodologies including, but not limited to, ultraviolet radiation absorbance, visible ray absorbance, fluorescence, refractive index, Raman or mass spectrometry, electrochemistry, and / or conductivity. Detection by fluorescence is preferred. Fluorescence detection using this module involves a microchip laser beam passing through the channels of the microchip 32 scans. The module can detect fluorescence using confocal optics.

The unit designed as a unit 62 can continue a sterile deionized unit 42 , a Siebgelpuffereinheit 44 and a microchannel reprocessing solution unit 46 include. As in 7 each of these three units is divided into halves, one half containing the fresh solutions and the other half containing waste solutions.

In operation, a sample is applied to the membrane of the sample card 28 deposited, and the card is in the device formed as a unit 62 used. In this example, the cells in the sample are lysed by the chemical reagents contained in the membrane. The cellular DNA or other sample components are then transferred to the membrane by heating with an oven 30 dried. At this point, the card can be removed and archived, or it can be used with continued processing. Alternatively, a Guthrie paper dry blood blot can be used to deposit the sample.

After drying the sample to the card, the card membranes are steam-heated, with sterile deionized from the unit 42 is used to extract the sample components in a small amount of liquid. As described above, the sealed ends of the capillaries become the array of sample delivery systems 6 heated to expel gas, moved to a position over the membranes and immersed in the liquid containing the sample. As the closed ends of the capillaries cool, the gas in the capillaries contracts and a sample is drawn into the capillaries. The capillaries are preferably preloaded with the reagents specific for the immunoassay or polynucleotide detection to be performed, as mentioned above.

After a suitable reaction time, the reaction products in the sample introduction channels of the microchip assembly 32 deposited. The array of sample delivery systems 6 typically moves to position the open end of the capillaries above the sample introduction channels. The sealed ends of the capillaries are then heated by a temperature controller so that gas trapped inside the capillary expands and forces a sample out of the capillary, as described above. After use, the array of sample delivery systems 6 and new sample delivery systems containing reagents for the next reaction of interest may be incorporated into the unitized device 62 be used. With the appropriate conditions and application can be a fresh array of sample delivery systems 6 to unit designed as a unit 62 are introduced by a coil is unrolled and cut to a desired length.

After deposition on the sample introduction channel of the microchip assembly, a pressure / vacuum unit manipulates 34 the pressure gradients in the interior of the sample introduction and separation channels of the microchip, whereby a portion of the sample, ie a sample plug, is moved into the separation channel. Application of pressure along the separation channel substantially results in the formation of a plug of a sample in the separation channel downstream of the compound, as previously discussed in US Pat 6 shown.

If the separation involves electrophoresis, after the sample plug has formed in the separation channel, the voltage generator becomes 36 used a voltage gradient axially along the separation channel of the microchip 32 to separate the components of the sample. A sieve medium is in preloaded the channels of the microchip. The buffer of the unit 44 is injected into the separation channels prior to formation of the sample plug.

When the samples reach the end of the separation channel, the optical scanning module scans 40 the separation channels to detect the presence of the detectable entities attached to the sample components by reacting with the chemical reagents in the array of sample delivery systems 6 are included. For polynucleotide identifications, the results of optical scanning are compared to data generated from genetic diagnosis experiments. These data are in the form of graphene intensity vs. time, which can be found in determining a match identity.

After running the analysis, pressure is released from the high pressure cartridge 38 used to apply a pressure at both ends of the separation channel to clean the channels of the microchip assembly. The channels are then retrieved from the unit using a reprocessing solution 46 recycled. The microchip assembly can then be reused in subsequent analyzes. Alternatively, the microchip assembly may be disposed of after use.

Compared with the use of conventional Volume control devices, such as Spraying and pumping, instructs thermally controlled sample delivery system of the invention less movable Parts that wear out can or require extensive maintenance. In addition, because the sample delivery system independently from an analytical tool, other benefits are realized. For example, can the sample delivery channels off inexpensive Be made of materials such. Kunststoffkapillarrohrmaterial, because of an optical quality or integrated electrodes are not required. Accordingly, a One-time use of a channel attractive, which is a cleaning step eliminate and / or eliminate cross-contamination.

In addition, because the channels typically not used directly in an analytical technique, can the channels be easily movable and a higher one Have degree of tolerance for positioning. That is, because the detection system of the analyzer typically stationary remains, must the optical alignment of a liquid detection capillary while the analysis of a plurality of samples for optimal accuracy once performed become. Next, if the sample delivery system is a chemical reagent contains and used to carry out a reaction, any particulates, which are present or during The reaction can be easily formed before the introduction of the Reaction products are filtered to an analyzer, whereby Clogging and / or inaccurate analysis can be prevented. These allow the above features that a simple and inexpensive Automation robotics is used.

Compared with a use of capillary action to deliver chemicals to mix and / or implement, shows a sample delivery system of Invention using pressure has several advantages. The surface of a Channel of a sample delivery system of the invention may be hydrophilic or hydrophobic be, as opposed to a capillary action surface, the a hydrophilic surface requires. Also with respect to the surface of the channel is the reproducibility a Probenlösungszumessung independently of surface characteristics and sample components. additionally allows that Sample delivery system of the invention has a direct control over the Metering samples and reagents and allowing a blistering routinely practiced becomes.

Compared with electroosmotic flow for delivery, mixing and / or reaction of chemicals, shows a sample delivery system of the invention that Pressure uses some of the same benefits compared to one Use of one discussed above Capillary action, i. surface characteristics and reproducibility of a solution metering. Furthermore the sample delivery system of the invention is typically in its solution composition to carry out analysis and / or chemical reactions. That Variables, such as pH, ionic strength, Buffer composition, chemical additives and solvents, are common unlimited, dependent from the special application. These variables are typical limited, so that an effective electro-osmotic flow occurs.

Therefore allows As described above, the present invention provides high-speed delivery of samples for execution from micro-sized reactions and / or analysis of biological Samples without the complexity, time, labor and biological risk exposure conventional Techniques. additional Aspects and embodiments of the invention are in view of the foregoing disclosure seen. Accordingly, the Scope of the invention limited only by the scope of the appended claims.

The The invention may be embodied in other specific forms.

Claims (10)

Thermally controlled sample delivery system comprising: a housing defining a channel, the channel including an open end and a closed end; and a temperature control device in thermal communication with the closed end of the channel to control the temperature of a thermally expandable fluid disposed in the channel to convey a sample into the channel; characterized in that it further comprises: an insulating partition in the channel, an outer insulator in thermal communication with the channel, or a combination of both. Thermally controlled sample delivery system according to claim 1: (a) where the housing a capillary comprises; and, optionally, the capillary surface is not is wettable; (b) further comprising an array of the channels; (C) wherein the channel has a plurality of open ends and a plurality includes introductory subchannels, connecting the open ends to the channel; (D) where the case limits a plurality of channels, the plurality of channels includes a common closed end; and or (e) at the thermally expandable fluid is a gas. Thermally controlled sample delivery system according to claim 1, further comprising at least one chemical reagent in the channel is arranged; and optionally: (a) where the chemical reagent immobilized in the canal; (b) where the chemical Reagent selected from the group is composed of an oligonucleotide, a peptide nucleic acid, a binding protein, an enzyme, a substrate, a ligand, a receptor, an antibody and an antigen exists; (c) further comprising a stabilizer for the chemical reagent; (d) where the chemical reagent has a detectable moiety or a chemical modifying moiety is marked; and or (e) where the chemical reagent has a first set of chemical reagents and a second set of includes chemical reagents and the first set of chemical Reagents from the second set of chemical reagents in the distance is arranged. Thermally controlled sample delivery system according to claim 1: (a) wherein the channel is a polymeric tubing or is molded in a polymer; (b) where the channel is through Enclose one in the longitudinal direction open channel formed into an organic or an inorganic Substrate, which is the case limited, etched or has been molded; (c) where the channel is not one constant internal diameter comprises; (d) further comprising Volume control device in connection with the channel; and or (e) further comprising a second temperature control device; and optionally, wherein the second temperature control device is a first Line and a second line limited where the temperature of each one independent of the lines is regulated; and further optionally, in which the second temperature control device is parallel moved to the channel. Scientific instrument, including thermal Controlled sample delivery system according to any one of claims 1-4. Scientific instrument according to claim 5, further comprising a computer in connection with the temperature control device to the Temperature control device to control. Method for dispensing a sample into a channel, comprising the steps: (a) providing a thermally controlled Delivery system comprising (i) a housing defining a channel, the channel comprising an open end and a closed end; and (ii) an insulating partition in the channel, an outer insulator in thermal communication with the channel or a combination of in which; and (iii) a temperature control device in thermal communication with the closed end of the canal; (b) exposure a sample on the open end of the channel to the sample in the Introduce channel; and (C) Contracting a thermally expandable fluid in the channel is arranged, using the temperature control device to the To encourage sample into the canal. A method according to claim 7: (i) in which introduction of the sample into the channel is accomplished by non-thermally controlled means; (ii) further comprising the step of: expanding the thermally expandable fluid; (iii) further comprising the step of: expanding the thermally expandable fluid following step (c) to eject a portion of the sample; (iv) wherein the channel comprises a capillary; (v) wherein the thermally expandable fluid comprises a gas; (vi) further comprising the step of: providing a second temperature control device in thermal communication with the channel to independently regulate a temperature; (vii) further comprising the step of: analyzing for a component in the sample; and / or (viii) further comprising the steps of: (a) providing a sample plug formation amount apparatus comprising: a housing defining a separation channel including a longitudinal axis and an insertion channel which is non-parallel to the longitudinal axis and forms a junction with the separation channel to convey a sample to be analyzed; and a pressure regulator in communication with the separation channel and the insertion channel; (b) applying a pressure differential to the introduction channel to deliver a portion of the sample in communication with the insertion channel to the junction; (c) applying a second pressure differential to the separation channel to convey another portion of the sample into the separation channel at the junction to form a sample plug; and the step of expanding the thermally expandable fluid following step (c) to eject a portion of the sample. The method of claim 7, wherein the channel continues comprises at least one chemical reagent disposed therein is; and optionally: (a) where the chemical reagent in the channel is immobilized; (b) where the chemical reagent selected from the group is composed of an oligonucleotide, a peptide nucleic acid, a binding protein, an enzyme, a substrate, a ligand, a receptor, an antibody and an antigen exists; (c) further comprising a stabilizer for the chemical reagent; (d) where the chemical reagent has a detectable moiety or a chemical modifying moiety is marked; and or (e) where the chemical reagent has a first set of chemical reagents and a second set of includes chemical reagents and the first set of chemical Reagents from the second set of chemical reagents in the distance is arranged. Method for dispensing samples into a channel, which are separated by a gas, the method comprising the steps includes: (a) providing a thermally controlled delivery system according to one of the claims 1-4; (B) Exposing the open end of the channel to a first sample; (C) Contracting a thermally expandable fluid in the channel is arranged, using the temperature control device, at least to convey a portion of the first sample into the channel; (d) exposure a gas on the open end of the channel; (e) Contract a thermally expandable fluid disposed in the channel, using the temperature control device to control a volume of the gas to promote the channel; (F) Exposing the open end of the channel to a second sample; (G) Contracting a thermally expandable fluid in the channel is arranged, using the temperature control device, at least to convey a portion of the second sample into the canal.